Enhanced anaerobic synthesis of rhamnolipids and emulsification of crude oil by genetically engineered Pseudomonas aeruginosa strains.

IF 4.9 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Feng Zhao, Yuting Wu, Lu Liu
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Abstract

Background: Pseudomonas aeruginosa is a facultative anaerobic bacterium widely distributed in oil reservoirs. Efficient production of rhamnolipids by P. aeruginosa in anoxic environments of oil reservoirs contributes to oil displacement. How to enhance the synthesis of rhamnolipids by P. aeruginosa under anaerobic conditions has become a new scientific question.

Results: Pseudomonas aeruginosa SG anaerobically synthesized much lower yield of rhamnolipids than that under aerobic conditions. The key genes rmlBDAC (m), rhlABRI (h) and rhlC (c) and their combination genes were overexpressed in P. aeruginosa. Among seven genetically engineered strains, P. aeruginosa SGhm anaerobically synthesized the highest yield of rhamnolipids, 1.34 g/L, which was about 4.5-fold higher than that of the wild-type strain SG (0.24 g/L). The results of Plackett-Burman (PB) design indicated that glycerol and nitrate were the nutrient factors that significantly influenced the anaerobic production of rhamnolipids by strain SGhm. Optimization of culture medium by response surface method improved the rhamnolipids production of strain SGhm to 1.54 g/L under anaerobic conditions. Pseudomonas aeruginosa SGhm emulsified oil in anaerobic tubes through anaerobic synthesis of rhamnolipids. The percentage of oil droplets formed with a diameter of 0-5 μm was 89.4%.

Conclusions: Genes rmlBDAC and rhlABRI were the best overexpression combination for improvement of rhamnolipids production in P. aeruginosa under anaerobic conditions. Overexpression of key genes and optimization of medium synergistically increase rhamnolipids production of P. aeruginosa under anaerobic conditions. Pseudomonas aeruginosa SGhm is promising for emulsification of crude oil through anaerobic synthesis of rhamnolipids. This study provided an excellent microbial strain resource and nutrients for in-situ production of rhamnolipids in anoxic environments of oil reservoirs.

转基因铜绿假单胞菌强化鼠李糖脂厌氧合成及原油乳化。
背景:铜绿假单胞菌是广泛分布于油藏中的兼性厌氧细菌。铜绿假单胞菌在油藏缺氧环境下高效产鼠李糖脂有助于驱油。如何在厌氧条件下提高铜绿假单胞菌对鼠李糖脂的合成成为一个新的科学问题。结果:铜绿假单胞菌SG厌氧合成鼠李糖脂的产率明显低于好氧条件下的产率。关键基因rmlBDAC (m)、rhlABRI (h)和rhlC (c)及其组合基因在铜绿假单胞菌中过表达。在7个基因工程菌株中,P. aeruginosa SGhm厌氧合成鼠李糖脂的产量最高,为1.34 g/L,比野生型菌株SG (0.24 g/L)高出约4.5倍。Plackett-Burman (PB)设计结果表明,甘油和硝酸盐是影响菌株SGhm厌氧生产鼠李糖脂的营养因子。通过响应面法优化培养基,使菌株SGhm在厌氧条件下的鼠李糖脂产量达到1.54 g/L。铜绿假单胞菌SGhm乳化油在厌氧管中厌氧合成鼠李糖脂。直径为0 ~ 5 μm的油滴形成率为89.4%。结论:在厌氧条件下,rmlBDAC和rhlABRI基因是提高铜绿假单胞菌鼠李糖脂产量的最佳过表达组合。在厌氧条件下,关键基因的过表达和培养基的优化协同提高了铜绿假单胞菌的鼠李糖脂产量。铜绿假单胞菌SGhm在厌氧合成鼠李糖脂乳化原油方面具有广阔的应用前景。该研究为油藏缺氧环境下原位生产鼠李糖脂提供了良好的微生物菌种资源和营养物质。
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来源期刊
Microbial Cell Factories
Microbial Cell Factories 工程技术-生物工程与应用微生物
CiteScore
9.30
自引率
4.70%
发文量
235
审稿时长
2.3 months
期刊介绍: Microbial Cell Factories is an open access peer-reviewed journal that covers any topic related to the development, use and investigation of microbial cells as producers of recombinant proteins and natural products, or as catalyzers of biological transformations of industrial interest. Microbial Cell Factories is the world leading, primary research journal fully focusing on Applied Microbiology. The journal is divided into the following editorial sections: -Metabolic engineering -Synthetic biology -Whole-cell biocatalysis -Microbial regulations -Recombinant protein production/bioprocessing -Production of natural compounds -Systems biology of cell factories -Microbial production processes -Cell-free systems
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